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43Выводы.
1. Методом ВЭЖХ определено содержание арбутина в плодах клюквы мелкоплодной.
2. Определены некоторые валидационные характеристики методики определения количественного содержания арбутина в плодах клюквы.
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STUDY OF THE QUALITATIVE COMPOSITION AND QUANTITATIVE CONTENT OF AMINO ACIDS IN SPINACH PLANT RAW MATERIAL OF KRASEN POLISSIA AND FANTASY
CULTIVARS
Petrovska U.,
PhD student of the Chemistry of Natural Compounds Department, National University of Pharmacy
Zhuravel I.,
Professor, Doctor of Pharmaceutical Sciences, Professor of the Chemistry of Natural Compounds Department, National University of Pharmacy
Gurieva I.
Associate Professor, PhD (PharmSc), Associate Professor of the Chemistry of Natural Compounds Department, National University of Pharmacy
Abstract
Spinach (Spinacia oleracea L.) is a herbaceous plant spread worldwide. Its leaves are consumed in food in both raw and cooked forms. Such popularity is due to its nutritional qualities and diverse chemical composition. Thus, according to the literature, spinach leaves contains phenolic compounds, vitamins, proteins, amino acids, fiber, chlorophylls, carotenoids, mineral compounds, etc. In addition, spinach leaves are used in folk medicine as an anti-inflammatory, antioxidant and general tonic agent.
Using the method of ion-exchange liquid column chromatography, 9 essential and 9 nonessential amino acids were identified in spinach leaves and seeds of Krasen Polissia and Fantasy cultivars. Glutamic acid prevailed by the quantitative content in the studied plant raw material of both cultivars. Its quantitative content in spinach leaves of Krasen Polissia cultivar comprised 2.903 mg/g, while in spinach leaves of Fantasy cultivar - 2.145 mg/g. Spinach seeds of Krasen Polissia cultivar contained 3.096 mg/g of glutamic acid, and spinach seeds of Fantasy cultivar - 2.154 mg/g.
The results obtained will be used for planning further phytochemical research of spinach plant raw material and creation of new herbal remedies on its basis.
Keywords: Spinacia oleracea, amino acids, ion-exchange chromatography.
Introduction.
Spinach (Spinacia oleracea L.) is a herbaceous plant of Spinacia genus, the Amaranth family (Amaranthaceae). Spinach originates from south-eastern Asia, from Ancient Persia, where spinach was first cultivated about 2000 ago. Nowadays it is one of the most widespread and consumed leafy green vegetable all over the world [12, p. 154; 19, p. 451.]. It is grown in America, Japan, countries of Western Europe and is widely cultivated in Ukraine.
The rosette of spinach leaves comprises of 15-20 leaves; it is dense, semi-submerged, 35-40 cm in diameter and 20-22 cm in height. The leaves are up to 20 cm long and 11-14 cm wide. They are petiolate, elongated-elliptic, the leaf margin is entire, vesicular. The shape
of the leaf blade is orbicular, elongated-elliptic, ovate or spear-shaped depending on the cultivar. Spinach is a dioecious plant [16, p. 77; 17, p. 1].
Spinach leaves are consumed fresh, boiled, baked, frozen, and are used in folk medicine as an anti-inflammatory, antioxidant and general tonic agent. Spinach leaves are used in anemia, for the treatment of digestive organs, upper respiratory tract disorders, after severe illnesses, etc. [2, p. 60; 9, p. 192]. Spinach are known to be used in cardiovascular diseases, Alzheimer's disease and diabetes mellitus [13, p. 37; 14, p. 80; 15, p. 106].
Such spectrum of application is explained by the diverse chemical composition of the plant. Its leaves contain phenolic compounds, proteins, organic acids,
saturated and unsaturated fatty acids, sugars, fiber, vitamins A, E, C, K, PP and B group, chlorophylls, carot-enoids, as well as mineral compounds, such as calcium, potassium, sodium, iron, magnesium, phosphorus and zinc [5, p. 532; 6, p. 17; 7, p. 3; 8, p. 2768]. However, it is worth mentioning that mature spinach leaves contains oxalic acid, which makes it contraindicated for people with urinary tract disorders, urolithiasis, nephritis, liver, gall bladder and pancreatic disorders [18, p. 1692].
Amino acids are primary synthesis compounds that are found in aerial parts of almost every plant, are synthesized from simple inorganic compounds and take part in the synthesis of proteins, flavonoids, steroidal compounds, complex carbohydrates, fats, vitamins and pigments. In addition, amino acids take part in the synthesis of enzymes, hormones, mediators, components of connective and muscle tissues, in blood formation processes, and are a part of coenzymes [3, p. 102; 10, p. 29; 11, p. 3997; 4, p. 42].
Amino acids are applied in medicine in the treatment of cardiovascular, nervous, digestive systems, for the immune system strengthening, normal functioning of endocrine glands, in atherosclerosis prophylaxis, and inhibiting the development of malignant neoplasms [1, p5]
The purpose of our work was the study of the qualitative composition and quantitative content of amino acids in spinach plant raw material of Krasen Polissia and Fantasy cultivars.
Materials and Methods.
The objects of the research were dried and crushed spinach leaves and seeds of Krasen Polissia and Fantasy cultivars. The plant material was grown on the fields of the Institute of Horticulture and Melon-growing of the National Academy of Agrarian Sciences of Ukraine in 2016-2017. The leaves were collected in the phase of maximal vegetative development, the seeds - during the fruiting stage.
Determination of qualitative composition and quantitative content of amino acids in the studied plant raw material was carried out by ion-exchange chromatography using the automatic amino acid analyzer AAA T-339M (Czech Republic) [1, p. 6; 3, p. 104; 20, p. 47].
Results and Discussion.
After the analysis carried out, 9 essential and 9 nonessential amino acids were detected in spinach plant raw material of both studied cultivars. Chromatograms of amino acids of the studied spinach plant raw material, obtained by the method of ion-exchange liquid column chromatography, are given in Figures 1,2.
Fig.1 Chromatogram of amino acids in spinach leaves of Krasen Polissia cultivar (A), in spinach seeds of
Krasen Polissia cultivar (B)
Fig.2 Chromatogram of amino acids in spinach leaves of Fantasy cultivar (A), in spinach seeds of Fantasy cultivar (B).
The qualitative composition and the quantitative content of amino acids in spinach leaves and seeds of Krasen
Polissia and Fantasy cultivars are given in the table.
№ Amino acid Content of an amino acid, mg/g
Spinach of Krasen Polissia cultivar Spinach of Fantasy cultivar
Leaves Seeds Leaves Seeds
Essential amino acids
1 Arginine 0.387 1.223 1.005 0.990
2 Valine 0.362 0.254 0.409 0.187
3 Histidine 0.423 0.305 0.304 0.221
4 Isoleucine 0.341 0.242 0.432 0.190
5 Leucine 0.953 0.628 1.175 0.487
6 Lysine 1.190 0.574 0.807 0.376
7 Methionine 0.190 0.154 0.242 0.117
8 Treonine 0.769 0.362 0.599 0.288
9 Phenylalanine 0.724 0.443 0.835 0.351
Nonessential amino acids
10 Alanine 0.960 0.601 1.038 0.441
11 Aspartic acid 1.787 1.095 1.404 0.926
12 Glycine 1.325 1.017 0.788 0.717
13 Glutamic acid 2.903 3.096 2.145 2.154
14 GABA 0.027 0.017 0.126 0.013
15 Proline 1.522 0.572 0.867 0.334
16 Serine 0.878 0.566 0.743 0.474
17 Tyrosine 0.546 0.270 0.614 0.180
18 Cystine 0.222 0.188 0.238 0.167
Total nonessential amino acids 5.339 4.185 5.808 3.207
Total essential amino acids 10.170 7.422 7.963 5.406
Total 15.509 11.607 13.771 8.613
Glutamic acid was found to be the dominating amino acid for the leaves and seeds of two studied cultivars. Its content in spinach leaves and seeds of Krasen Polissia cultivar comprised 2,903 mg/g and 3,096 mg/g respectively. The content of this amino acid for the spinach plant raw material of Fantasy cultivar was somewhat lower- 2,145 mg/g in the leaves and 2,154 mg/g in the seeds.
Aspartic acid was found in the leaves of both spinach cultivars. The content of this amino acid for spinach leaves of Krasen Polissia cultivar amounted to 1,787 mg/g and for spinach leaves of Fantasy cultivar - 1,404 mg/g.
The amount of arginine in spinach seeds of Krasen Polissia cultivar was 1,223 mg/g, while in spinach seeds of Fantasy cultivar - 0,990 mg/g. The accumulation of aspartic acid and glycine was characterized by somewhat lower numbers. The content of aspartic acid in spinach seeds of Krasen Polissia cultivar amounted to 1,095 mg/g, and in spinach seeds of Fantasy cultivar - 0,926 mg/g. The quantitative content of glycine in the seeds of Krasen Polissia cultivar was 1,017 mg/g, and in the seeds of Fantasy cultivar - 0,717 mg/g.
The quantitative content of proline was also determined in spinach leaves of Krasen Polissia cultivar. Its amount was 1,522 mg/g. The nonessential amino acid lysine accumulated in spinach leaves of Krasen Polissia cultivar in the amount of 1,190 mg/g. The content of other amino acids was lower than 1,000 mg/g.
The content of leucine in spinach leaves of Fantasy cultivar was 1,175 mg/g. Alanine and the essential amino acid arginine accumulated in somewhat lower amount in this plant raw material (1,038 mg/g and
1,005 mg/g respectively). The content of other amino acids was lower than 1,000 mg/g.
Conclusions.
The qualitative composition and the quantitative content of amino acids were determined in spinach leaves and seeds of Krasen Polissia and Fantasy culti-vars. As a result, 9 essential and 9 nonessential amino acids were detected. Glutamic acid was the dominating amino acid in the plant raw material of both cultivars. The results obtained will be further used for planning phytochemical research of spinach plant raw material and creation of new herbal remedies on its basis.
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